An official website of the United States government
The live visualization with fast kinetics of the interaction between cells has been long term challenges because of the lack of efficient stimulation methods. We propose an approach to achieve single cell resolution stimulation and FRET-base calcium live cell imaging to visualize fast kinetics of calcium transport between physically connect neighboring cells. Chemical stimulation stimulates cells within a dish at the same time and is not suitable for the study of cell-cell interaction. We replaced chemical stimulation with ultrasound-based mechanical stimulation approach to provide precise spatiotemporal resolution. To achieve this, we integrated 3D translation stages and epi-fluorescence microscope and a developed 150 MHz high frequency ultrasound with f number of 1 and aperture size of 1 mm. The 150 MHz transducer can focus within 10 micrometers in diameter and directly stimulate cells by disturbing cell plasma membranes without microbubbles. High frequency stimulation was used to introduce calcium ions into cytoplasm of cells. Results demonstrate calcium transport between cells, visualized by FRET calcium biosensor after only one cell was stimulated by the developed high frequency ultrasonic transducer.
more »
« less
RAPID FABRICATION OF MULTIFUNCTIONAL MICROCAPILLARY FOR FOUR DIMENSIONAL SINGLE CELL MANIPULATION
We report a novel manufacturing approach to fabricate liquid metal-based, multifunctional microcapillary pipettes able to provide electrodes with high electrical conductivity for high frequency electrical stimulation and measurement. Four-dimensional single cell manipulation has been realized by applying multi-frequency, multi-amplitude, and multi-phase electrical signals to the microelectrodes near the pipette tip to create a 3D dielectrophoretic trap and 1D electrorotation simultaneously. Functions such as single cell trapping, transferring, patterning, and rotation have been accomplished. Cell viability and multi-day proliferation characterization has confirmed the biocompatibility of this approach. This is a simple, low cost, and fast fabrication approach that requires no cleanroom and photolithography to manufacture 3D microelectrodes and microchannels accessible to a wide user base for broad applications.
more »
« less
- Award ID(s):
- 1711507
- PAR ID:
- 10063982
- Date Published:
- Journal Name:
- Proceedings, IEEE micro electro mechanical systems
- ISSN:
- 1084-6999
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
More Like this
-
-
null (Ed.)Microfabrication and assembly of a Three-Dimensional Microneedle Electrode Array (3D MEA) based on a glass-stainless steel platform is demonstrated involving the utilization of non-traditional “Makerspace Microfabrication” techniques featuring cost-effective, rapid fabrication and an assorted biocompatible material palette. The stainless steel microneedle electrode array was realized by planar laser micromachining and out-of-plane transitioning to have a 3D configuration with perpendicular transition angles. The 3D MEA chip is bonded onto a glass die with metal traces routed to the periphery of the chip for electrical interfacing. Confined precision drop casting (CPDC) of PDMS is used to define an insulation layer and realize the 3D microelectrodes. The use of glass as a substrate offers optical clarity allowing for simultaneous optical and electrical probing of electrogenic cells. Additionally, an interconnect using 3D printing and conductive ink casting has been developed which allows metal traces on the glass chip to be transitioned to the bottomside of the device for interfacing with commercial data acquisition/analysis equipment. The 3D MEAs demonstrate an average impedance/phase of ∼13.3 kΩ/−12.1° at 1 kHz respectively, and an average 4.2 μV noise. Lastly, electrophysiological activity from an immortal cardiomyocyte cell line was recorded using the 3D MEA demonstrating end to end device development.more » « less
-
Abstract Bioelectronic devices that allow simultaneous accurate monitoring and control of the spatiotemporal patterns of cardiac activity provide an effective means to understand the mechanisms and optimize therapeutic strategies for heart disease. Optogenetics is a promising technology for cardiac research due to its advantages such as cell‐type selectivity and high space‐time resolution, but its efficacy is limited by the insufficient number of modulation channels and lack of simultaneous spatiotemporal mapping capabilities in current implantable cardiac optogenetics tools available for in vivo investigations. Here, soft implantable electro‐optical cardiac devices integrating multilayered highly uniform arrays of transparent microelectrodes and multicolor light‐emitting diodes in thin, flexible platforms are designed for mechanically compliant high‐content high‐precision electrical mapping and single‐/multi‐site optogenetics and electrical stimulation without light‐induced artifacts. Systematic benchtop characterizations, together with ex vivo and in vivo evaluations on healthy and diseased small animal hearts and human cardiac slices demonstrate their functionalities in real‐time spatiotemporal mapping and control of cardiac rhythm and function, with broad applications in basic and ultimately clinical cardiology.more » « less
-
Many recent efforts in the diagnostic field address the accessibility of cancer diagnosis. Typical histological staining methods identify cancer cells visually by a larger nucleus with more condensed chromatin. Machine learning (ML) has been incorporated into image analysis for improving this process. Recently, impedance spectrometers have been shown to generate all-inclusive lab-on-a-chip platforms to detect nucleus abnormities. In this paper, a wideband electrical sensor and data analysis paradigm that can identify nuclear changes shows the realization of a single-cell microfluidic device to detect nuclei of altered sizes. To model cells of altered nucleus, Jurkat cells were treated to enlarge or shrink their nucleus followed by broadband sensing to obtain the S-parameters of single cells. The ability to deduce important frequencies associated with nucleus size is demonstrated and used to improve classification models in both binary and multiclass scenarios, despite a heterogeneous and overlapping cell population. The important frequency features match those predicted in a double-shell circuit model published in prior work, demonstrating a coherent new analytical technique for electrical data analysis. The electrical sensing platform assisted by ML with impressive accuracy of cell classification looks forward to a label-free and flexible approach to cancer diagnosis.more » « less
-
Abstract Despite ongoing advances in our understanding of local single-cellular and network-level activity of neuronal populations in the human brain, extraordinarily little is known about their “intermediate” microscale local circuit dynamics. Here, we utilized ultra-high-density microelectrode arrays and a rare opportunity to perform intracranial recordings across multiple cortical areas in human participants to discover three distinct classes of cortical activity that are not locked to ongoing natural brain rhythmic activity. The first included fast waveforms similar to extracellular single-unit activity. The other two types were discrete events with slower waveform dynamics and were found preferentially in upper cortical layers. These second and third types were also observed in rodents, nonhuman primates, and semi-chronic recordings from humans via laminar and Utah array microelectrodes. The rates of all three events were selectively modulated by auditory and electrical stimuli, pharmacological manipulation, and cold saline application and had small causal co-occurrences. These results suggest that the proper combination of high-resolution microelectrodes and analytic techniques can capture neuronal dynamics that lay between somatic action potentials and aggregate population activity. Understanding intermediate microscale dynamics in relation to single-cell and network dynamics may reveal important details about activity in the full cortical circuit.more » « less
- Free Publicly Accessible Full Text
-
Accepted Manuscript1.0
- Conference Paper:
- The DOI is not currently available.
